A manual of rice seed health testing - IRRI books - International Rice ...
A manual of rice seed health testing - IRRI books - International Rice ...
A manual of rice seed health testing - IRRI books - International Rice ...
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The blotter test, an indirect<br />
method <strong>of</strong> major importance, requires<br />
near-ultra violet (NUV) light<br />
incubation chambers equipped with<br />
timers to regulate NUV for 12 h, followed<br />
by 12 h darkness, daily. The<br />
incubation chamber temperature for<br />
the standard blotter test is 20±2 °C.<br />
Some NUV chambers are constructed<br />
inside a walk-in room that<br />
is air-conditioned to maintain the<br />
recommended temperature. The<br />
NUV light source should be a 320-<br />
400 nm lamp, preferably Philips TL<br />
40W/08, GE F 40 BL, or equivalent.<br />
A freezing blotter may be used to<br />
kill the germinating embryo or <strong>seed</strong>ling<br />
when germination is so vigorous<br />
that <strong>seed</strong>ling growth elevates the<br />
petri dish cover and the blotter becomes<br />
dry. Freezing is accomplished<br />
on the second or third day <strong>of</strong> incubation<br />
by placing the <strong>seed</strong>ed plates in a<br />
freezer for 24 h and then returning<br />
them to NUV incubation. Plastic<br />
petri dishes are recommended as<br />
they allow better NUV light penetration<br />
than do glass dishes.<br />
Agar for fungi test should be incubated<br />
at 28±1 °C in NUV light/<br />
darkness, and that for bacteria at<br />
28 °C. This can be done in standard<br />
laboratory-top incubators.<br />
For the growing-on test, container<br />
covers for the medium (substrate)<br />
and test <strong>seed</strong>s should be transparent<br />
to admit light and to facilitate visual<br />
examination without removing the<br />
covers. Transparent plastic bags supported<br />
by sticks or ready-made hard<br />
transparent covers used in germination<br />
tests are available.<br />
Table 3.1. Laboratory equipment, instruments, and glassware used for <strong>seed</strong> <strong>health</strong> <strong>testing</strong>. a<br />
Seed sampling<br />
Stick or sleeve trier (Nobbe trier. Fig. 4.1)<br />
Conical divider (Boerner type, Fig. 4.3)<br />
Soil divider (Riffle type, Fig. 4.4)<br />
Centrifugal divider (Gamet type. Fig. 4.5)<br />
Magnifying lens (Fig. 3.3)<br />
Seed <strong>health</strong> <strong>testing</strong><br />
Essential<br />
Autoclave<br />
Sterilizing oven<br />
Hot air<br />
Refrigerator/freezer<br />
Clean bench or laminar flow hood<br />
Centrifuge (up to 20,000 rpm)<br />
Centrifuge (table top model - 3,000-5,000 rpm)<br />
Automatic shaker or rotary shaker (adjustable speed)<br />
Distilling apparatus<br />
Incubation chambers (25-32 °C)<br />
NUV incubator (21 °C) with automatic time switch for lamps<br />
Lighted incubator (25-32 °C)<br />
Stereobinocular microscope - magnification up to 50X<br />
Compound microscope - magnification up to 1000X<br />
Mineral light lamp-UV<br />
Hand lens<br />
Haemocytometer<br />
Tally counter<br />
Mortar and pestle<br />
Electric mixer<br />
Vortex mixer<br />
Hot water bath<br />
pH meter<br />
Alcohol lamp<br />
Optional<br />
Growth chamber<br />
Hot plate<br />
Filtration apparatus<br />
Gas burner<br />
Electrostatic air cleaner<br />
Phase-contrast microscope<br />
Glassware and other laboratory supplies<br />
Petri dishes - glass, plastic<br />
Test tubes - assorted sizes<br />
Beakers - assorted sizes<br />
Syringes - assorted sizes<br />
Erlenmeyer flasks - assorted sizes<br />
Atomizer or sprayer<br />
Baermann funnel set-up (Fig. 8.1)<br />
Funnels - glass, plastic<br />
Graduated cylinders - 50, 100, 500, 1000 ml<br />
Pipettes - assorted sizes<br />
Germination trays with transparent covers<br />
Glass rods, L-shaped<br />
Microscope slides/cover glasses<br />
Filter paper<br />
Blotters, round, 8.5 cm diameter<br />
Wire loop (transfer loop)<br />
Inoculating needle<br />
Plastic bags - different sizes<br />
Mineral oil or paraffin<br />
Marking pens for glass<br />
Indelible pencils<br />
Paper towels<br />
Working box containing dissecting needles, forceps, scissors, scalpels,<br />
transfer needles, etc.<br />
a Sources: IRRl Seed Health Unit and Langerak et al (1988).<br />
14 <strong>Rice</strong> <strong>seed</strong> <strong>health</strong> <strong>testing</strong> <strong>manual</strong>